Accessory enzymes of hypercellulolytic Penicillium funiculosum facilitate complete saccharification of sugarcane bagasse.
Accessory enzymes of hypercellulolytic Penicillium funiculosum facilitate complete saccharification of sugarcane bagasse.
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DOI：
10.1186/s13068-021-02020-x
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发表时间：
2021-08-26
影响因子：
6.3
通讯作者：
Yazdani SS
中科院分区：
文献类型：
--
作者：
Ogunyewo OA;Upadhyay P;Rajacharya GH;Okereke OE;Faas L;Gómez LD;McQueen-Mason SJ;Yazdani SS
甘蔗渣是生产第二代生物乙醇的丰富原料。这种复杂的生物质需要一系列碳水化合物活性酶(CAZymes)，主要来自丝状真菌，以将其分解为单体糖，以生产增值燃料和化学品。在这项研究中，我们评估了一株分解代谢抑制因子缺陷菌株PfMig188分泌组中的蛋白质对SCB诱导的响应，并研究了它们在SCB糖化中的作用。开发了一种系统的方法来培养这种真菌，目的是生产和了解为SCB糖化量身定做的酶阵列。为了实现这一点，真菌生长在添加不同浓度的预处理SCB(0-45微克/L)的培养基中。采用酶活性测定和LC-MS/MS联用技术对分泌蛋白进行鉴定。在PfMig188的分泌组中共鉴定出280个蛋白质，其中46%被明确鉴定为CAZymes。结果表明，在高达15 g/L的条件下，半纤维素酶和细胞壁修饰酶的分泌依次增加，包括内切β-1，3(4)-葡聚糖酶(GH16)、内切-α-1，3-葡聚糖酶(GH71)、木聚糖酶(GH30)、β-木糖苷酶(GH5)、β-1，3-半乳糖苷酶(GH43)和角质酶(CE5)。 木糖苷酶和角质酶活性分别提高122%和60%。混合连接葡聚糖的活性也增加了36%。这些酶在分泌体中的诱导使糖化性能提高到98%(~ 比对照提高20%)，表明它们与核心纤维素酶在进入球状芽孢杆菌的顽固区方面具有协同作用。我们的发现提供了对PfMig188降解SCB等复杂生物量的酶系统的洞察，并强调了在培养基中添加SCB以优化蔗渣糖化专用酶的分泌的重要性。网上版载有补充材料，可在10.1186/s13068-021-02020-x查阅。
Sugarcane bagasse (SCB) is an abundant feedstock for second-generation bioethanol production. This complex biomass requires an array of carbohydrate active enzymes (CAZymes), mostly from filamentous fungi, for its deconstruction to monomeric sugars for the production of value-added fuels and chemicals. In this study, we evaluated the repertoire of proteins in the secretome of a catabolite repressor-deficient strain of Penicillium funiculosum, PfMig188, in response to SCB induction and examined their role in the saccharification of SCB. A systematic approach was developed for the cultivation of the fungus with the aim of producing and understanding arrays of enzymes tailored for saccharification of SCB. To achieve this, the fungus was grown in media supplemented with different concentrations of pretreated SCB (0–45 g/L). The profile of secreted proteins was characterized by enzyme activity assays and liquid chromatography–tandem mass spectrometry (LC–MS/MS). A total of 280 proteins were identified in the secretome of PfMig188, 46% of them being clearly identified as CAZymes. Modulation of the cultivation media with SCB up to 15 g/L led to sequential enhancement in the secretion of hemicellulases and cell wall-modifying enzymes, including endo-β-1,3(4)-glucanase (GH16), endo-α-1,3-glucanase (GH71), xylanase (GH30), β-xylosidase (GH5), β-1,3-galactosidase (GH43) and cutinase (CE5). There was ~ 122% and 60% increases in β-xylosidase and cutinase activities, respectively. There was also a 36% increase in activities towards mixed-linked glucans. Induction of these enzymes in the secretome improved the saccharification performance to 98% (~ 20% increase over control), suggesting their synergy with core cellulases in accessing the recalcitrant region of SCB. Our findings provide an insight into the enzyme system of PfMig188 for degradation of complex biomass such as SCB and highlight the importance of adding SCB to the culture medium to optimize the secretion of enzymes specific for the saccharification of sugarcane bagasse. The online version contains supplementary material available at 10.1186/s13068-021-02020-x.
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影响因子：
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